The H9c2 cells are seeded in the 96 well at a density of 1×10⁵ cells/well. The cells are treated with different concentrations of Trigonelline chloride (TG) (25 to 150 μM) and hydrogen peroxide (25 to 125 μM). It is incubated at 37°C in 5% CO₂ incubator for 24 h and 6 h respectively and then the culture is treated with the water soluble tetrazolium (WST) reagent incubated for 2 h to 4 h. The living cells absorb the WST then it is converted into an orange colour product. Then, the intensity of colour is measured at 450 nm using spectra count ELISA reader. For cardio protective activity, the cells are seeded and separated into six groups: control, H₂O₂ alone, the rest of groups are initially exposed to different concentration (25 to 125 μM) of Trigonelline chloride for 48 hours. Then, 100 μM of H₂O₂ is added and incubated for 4 hours, after, read the absorbance at 450 nm for cell viability assay^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Three-month-old female Wistar rats are used in this study. The animals are divided into five groups (n=10): Control rats, Streptozotocin-treated control rats, Streptozotocin-treated rats receiving Trigonelline chloride (50 mg/kg p.o. daily), Nicotinamide/streptozotocin-treated control rats, and Nicotinamide/streptozotocin-treated rats receiving Trigonelline chloride (50 mg/kg p.o. daily). Administration of Trigonelline chloride starts two weeks after streptozotocin and lasts four weeks. Trigonelline chloride is administered once daily by a stomach tube. All control rats receive tap water (the vehicle) at the same volume of 2 mL/kg p.o. The four-week period of Trigonelline chloride administration is long enough to demonstrate skeletal effects of Trigonelline chloride and other compounds of plant origin in rats. The rats are fasted overnight after the last Trigonelline chloride or vehicle administration. The next day, the blood glucose level is measured and the rats are anesthetized with ketamine and xylazine, and then sacrificed by cardiac exsanguination^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Ilavenil S, et al. Trigonelline protects the cardiocyte from hydrogen peroxide induced apoptosis in H9c2 cells. Asian Pac J Trop Med. 2015 Apr;8(4):263-8.

  [2]. Joanna Folwarczna, et al. Effects of Trigonelline, an Alkaloid Present in Coffee, on Diabetes-Induced Disorders in the Rat Skeletal System. Nutrients. 2016 Mar; 8(3): 133.

173.60

C₇H₈ClNO₂

6138-41-6

C[N+]1=CC(C(O)=O)=CC=C1.[Cl-]

Room temperature in continental US; may vary elsewhere